Trisubstituted acridine derivative containing piperazine group as well as preparation method and application of trisubstituted acridine derivative

Abstract

The invention belongs to the technical field of acridine derivatives, and provides a trisubstituted acridine derivative containing a piperazine group as well as a preparation method and application of the trisubstituted acridine derivative. The structural formula of the trisubstituted acridine derivative containing the piperazine group disclosed by the invention is # imgabs0. The trisubstituted acridine derivative containing the piperazine group disclosed by the invention has excellent selectivity and stability on a four-helix structure (G4-DNA), and also has broad-spectrum anti-tumor activity and anti-inflammatory activity; the traditional Chinese medicine composition contains but is not limited to ovarian cancer, breast cancer, colorectal cancer, lung cancer, liver cancer, brain glioma and the like, and has relatively low toxic and side effects on normal cells and tissues.

Description

Technical Field

[0001] The present invention relates to the technical field of acridine derivatives, and particularly relates to a trisubstituted acridine derivative containing a piperazine group, and a preparation method and application thereof. Background Art

[0002] At present, cancer is a disease with a relatively high mortality rate, the incidence rate is continuously increasing, and the patients show a trend of getting younger. Drug therapy is the main treatment strategy for patients with medium and advanced cancer. Although certain successes have been achieved in the treatment of some cancers, there are still huge limitations. For example, drugs have serious toxic and side effects on normal cells, cancer cells develop drug resistance to drugs, and the unique heterogeneity of cancer cells. Therefore, finding effective anti-cancer drugs has become an urgent task in public health and medical research. Inflammation is a complex biological response caused by infection, trauma, immune disorders or other stimuli, and usually manifests as redness, swelling, heat, pain, dysfunction, etc. Currently, anti-inflammatory drugs generally have relatively large toxic and side effects. Most importantly, there is a close connection between the pathology of inflammation and cancer. For example, inflammation may promote the occurrence and development of cancer, and abnormal inflammatory responses are bound to occur during the pathological progression of cancer. Therefore, developing drugs with both anti-cancer and anti-inflammatory dual activities has a broader prospect than single anti-cancer drugs.

[0003] Acridine is a nitrogen-containing heterocyclic compound with a tricyclic aromatic structure. Acridine derivatives obtained by modifying different chemical groups can exhibit various biological activities. Due to its strong affinity for nucleic acids, especially the interaction with DNA and RNA, acridine derivatives have become potential anti-tumor drug components. At present, there are still certain defects and challenges for acridine derivatives as anti-tumor drugs, mainly manifested in their lack of selectivity for tumor cells and normal cells, and having serious toxic and side effects. The four-stranded structure (G4-DNA) can be significantly distinguished from the conventional double-stranded DNA structure, and G4-DNA is widely present in the promoter regions of proto-oncogenes, which is a potential new anti-tumor drug target. Acridine derivatives obtained by modifying different chemical groups are expected to exhibit high activity only against G4-DNA, effectively improving the selectivity of acridine derivatives for tumor cells and reducing their toxic and side effects. However, there has not been much research on whether acridine derivatives have anti-inflammatory activity.

[0004] Therefore, it is of great significance to study trisubstituted acridine derivatives with high anti-tumor activity, selectivity, low toxic and side effects and anti-inflammatory activity targeting G4-DNA. Summary of the Invention

[0005] The object of the present invention is to provide a trisubstituted acridine derivative containing a piperazine group, a preparation method thereof and an application thereof in view of the deficiencies of the prior art.

[0006] In order to achieve the above object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides a trisubstituted acridine derivative containing a piperazine group, and the structural formula of the trisubstituted acridine derivative containing a piperazine group is:

[0008]

[0009] The present invention also provides a preparation method of the trisubstituted acridine derivative containing a piperazine group, which comprises the following steps:

[0010] Mix a trisubstituted acridine intermediate, N-methylpiperazine, potassium iodide, potassium carbonate and absolute ethanol, and carry out a nucleophilic substitution reaction to obtain a trisubstituted acridine derivative containing a piperazine group.

[0011] Preferably, the structural formula of the trisubstituted acridine intermediate is:

[0012]

[0013] Preferably, the molar ratio of the trisubstituted acridine intermediate, N-methylpiperazine, potassium iodide and potassium carbonate is 1:4-6:0.4-0.6:0.4-0.6, and the molar volume ratio of the trisubstituted acridine intermediate and absolute ethanol is 1 mmol:13-18 mL.

[0014] Preferably, the temperature of the nucleophilic substitution reaction is 80-100 °C, and the time of the nucleophilic substitution reaction is 1-3 h.

[0015] Preferably, the nucleophilic substitution reaction is carried out under stirring conditions, and the stirring speed is 400-600 rpm.

[0016] The present invention also provides an application of the trisubstituted acridine derivative containing a piperazine group in the preparation of anti-tumor drugs and anti-inflammatory drugs.

[0017] The beneficial effects of the present invention:

[0018] The trisubstituted acridine derivative containing a piperazine group of the present invention has excellent selectivity and stability for the four-stranded structure (G4-DNA), and at the same time has broad-spectrum anti-tumor activity and anti-inflammatory activity, including but not limited to ovarian cancer, breast cancer, colorectal cancer, lung cancer, liver cancer, glioblastoma, etc., and has low toxicity and side effects on normal cells and tissues. Description of the Drawings

[0019] Figure 11H NMR nuclear magnetic resonance spectrum of the trisubstituted acridine derivative containing a piperazine group prepared in Example 1; 1 HNMR nuclear magnetic resonance spectrum;

[0020] Figure 2 Inhibitory activity of the trisubstituted acridine derivative containing a piperazine group prepared in Example 1 against tumor cells;

[0021] Figure 3 Inhibitory activity of the trisubstituted acridine derivative containing a piperazine group prepared in Example 1 against inflammatory factors. Detailed implementation manners

[0022] The present invention provides a trisubstituted acridine derivative containing a piperazine group, and the structural formula of the trisubstituted acridine derivative containing a piperazine group is:

[0023]

[0024] The present invention also provides a preparation method of the trisubstituted acridine derivative containing a piperazine group, comprising the following steps:

[0025] Mix a trisubstituted acridine intermediate, N-methylpiperazine, potassium iodide, potassium carbonate and absolute ethanol, and carry out a nucleophilic substitution reaction to obtain the trisubstituted acridine derivative containing a piperazine group.

[0026] In the present invention, the structural formula of the trisubstituted acridine intermediate is preferably:

[0027]

[0028] In the present invention, the molar ratio of the trisubstituted acridine intermediate, N-methylpiperazine, potassium iodide and potassium carbonate is preferably 1:4-6:0.4-0.6:0.4-0.6, more preferably 1:4.5-5.5:0.45-0.55:0.45-0.55, and even more preferably 1:5:0.5:0.5; the molar volume ratio of the trisubstituted acridine intermediate and absolute ethanol is preferably 1 mmol:13-18 mL, more preferably 1 mmol:14-17 mL, and even more preferably 1 mmol:15-16 mL.

[0029] In the present invention, the temperature of the nucleophilic substitution reaction is preferably 80-100 °C, more preferably 85-95 °C, and even more preferably 90 °C; the time of the nucleophilic substitution reaction is preferably 1-3 h, more preferably 1.5-2.5 h, and even more preferably 2 h.

[0030] In the present invention, the nucleophilic substitution reaction is preferably carried out under stirring conditions, and the stirring speed is preferably 400-600 rpm, more preferably 450-550 rpm, and even more preferably 500 rpm.

[0031] In the present invention, it is preferred that the mixing first mixes a trisubstituted acridine intermediate, potassium iodide and potassium carbonate, and then sequentially adds absolute ethanol and N-methylpiperazine.

[0032] In the present invention, after the nucleophilic substitution reaction is completed, it is preferred to sequentially perform a quenching reaction, distillation under reduced pressure, extraction to obtain an organic layer, and then sequentially perform drying, distillation under reduced pressure, and silica gel column chromatography on the organic layer to obtain a trisubstituted acridine derivative containing a piperazine group; the reagent used for the quenching reaction is water, and the volume ratio of water to absolute ethanol is preferably 20-30:1, more preferably 25:1; the temperature of distillation under reduced pressure is independently preferably 45-55 °C, more preferably 50 °C; the pressure of distillation under reduced pressure is independently preferably 0.05-0.15 MPa, more preferably 0.1 MPa; the time of distillation under reduced pressure is independently preferably 20-30 min, more preferably 25 min; the reagents used for the extraction are preferably dichloromethane and saturated sodium chloride solution, and the volume ratio of dichloromethane to saturated sodium chloride solution is preferably 1-2:1, more preferably 1.5:1; the number of extractions is preferably 2-4 times, more preferably 3 times; the reagent used for the drying is preferably anhydrous magnesium sulfate; the reagents used for the silica gel column chromatography are preferably dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is preferably 10-30:1, more preferably 20:1.

[0033] In the present invention, the trisubstituted acridine derivative containing a piperazine group is N,N'-(9-((4-(dimethylamino)phenyl)amino)acridine-3,6-diyl)bis(3-(4-methylpiperazin-1-yl)propanamide).

[0034] In the present invention, the preparation method of the trisubstituted acridine intermediate preferably comprises the following steps:

[0035] 1) Condensing 2-chloro-4-aminobenzoic acid and acetic anhydride to obtain a condensation intermediate;

[0036] 2) Mixing the condensation intermediate, m-aminophenetidine, copper powder, cuprous oxide, potassium carbonate and ethylene glycol monoethyl ether, and performing an Ullmann reaction to obtain a benzoic acid intermediate;

[0037] 3) Performing a cyclization reaction on the benzoic acid intermediate and sulfuric acid to obtain an acridone intermediate 1;

[0038] 4) Condensing the acridone intermediate 1 and butyric anhydride to obtain an acridone intermediate 2;

[0039] 5) Performing an electrophilic substitution reaction on the acridone intermediate 2 and phosphorus oxychloride to obtain an acridine intermediate;

[0040] 6) Mixing the acridine intermediate, N,N-dimethyl-p-phenylenediamine and phenol, and performing a nucleophilic substitution reaction to obtain a trisubstituted acridine intermediate 1;

[0041] 7) Hydrolyze the trisubstituted acridine intermediate 1 and sulfuric acid to obtain the trisubstituted acridine intermediate 2;

[0042] 8) Mix the trisubstituted acridine intermediate 2, 3-chloropropionyl chloride, triethylamine, and N,N-dimethylformamide, and carry out a nucleophilic addition reaction to obtain the trisubstituted acridine intermediate.

[0043] In the present invention, the molar volume ratio of the 2-chloro-4-aminobenzoic acid and acetic anhydride in step 1) is preferably 110-120 mmol:30 mL, more preferably 112-118 mmol:30 mL, and even more preferably 116.6 mmol:30 mL; the temperature of the condensation reaction is preferably 90-110 °C, more preferably 95-105 °C, and even more preferably 100 °C; the time of the condensation reaction is preferably 20-40 min, more preferably 25-35 min, and even more preferably 30 min.

[0044] In the present invention, the condensation reaction in step 1) is preferably carried out under stirring conditions, and the stirring speed is preferably 700-900 rpm, more preferably 800 rpm.

[0045] In the present invention, after the condensation reaction in step 1) is completed, it is preferably to first add dichloromethane to precipitate, and then filter and wash to obtain the condensation intermediate; the volume ratio of dichloromethane to acetic anhydride is preferably 2-4:1, more preferably 3:1; the reagent used for washing is preferably dichloromethane, and the number of washing times is preferably 2-4 times, more preferably 3 times. The purpose of filtration is to remove excess acetic anhydride, by-product acetic acid, and dichloromethane.

[0046] In the present invention, the condensation intermediate in step 1) is 4-(acetylamino)-2-chlorobenzoic acid, and the structural formula is:

[0047] In the present invention, the molar ratio of the condensation intermediate, m-aminoacetanilide, copper powder, cuprous oxide and potassium carbonate described in step 2) is preferably 1:0.8 - 1.2:0.08 - 0.12:0.08 - 0.12:0.2 - 0.8, more preferably 1:0.9 - 1.1:0.09 - 0.11:0.09 - 0.11:0.3 - 0.6, and even more preferably 1:1.0:0.098:0.092:0.5; the molar volume ratio of the condensation intermediate and ethylene glycol monoethyl ether is preferably 1 mmol:1 - 2.5 mL, more preferably 1 mmol:1.3 - 2 mL, and even more preferably 1 mmol:1.7 mL; the temperature of the Ullmann reaction is preferably 125 - 145 °C, more preferably 130 - 140 °C, and even more preferably 135 °C; the time of the Ullmann reaction is preferably 4 - 6 h, more preferably 4.5 - 5.5 h, and even more preferably 5 h.

[0048] In the present invention, the Ullmann reaction described in step 2) is preferably carried out under stirring conditions, and the stirring speed is preferably 400 - 600 rpm, more preferably 500 rpm.

[0049] In the present invention, after the Ullmann reaction described in step 2) is completed, a quenching reaction, filtration, recrystallization and drying are preferably carried out in sequence to obtain a benzoic acid intermediate; the reagent used for the quenching reaction is water, and the volume ratio of water to ethylene glycol monoethyl ether is preferably 3 - 4:1, more preferably 3.75:1; the recrystallization is preferably carried out by acidifying the filtrate obtained by filtration with hydrochloric acid, and the volume ratio of HCl to water in the hydrochloric acid is preferably 2.5 - 3.5:1, more preferably 3:1, and the acidification ends when the pH value of the filtrate is 2 - 3; the drying temperature is preferably 40 - 60 °C, more preferably 50 °C; the drying time is preferably 20 - 28 h, more preferably 24 h. The purpose of filtration is to remove the catalyst copper powder and cuprous oxide.

[0050] In the present invention, the benzoic acid intermediate described in step 2) is 4-(acetamido)-2-(3-(acetamido)phenylamino)benzoic acid, and the structural formula is:

[0051] In the present invention, the molar volume ratio of the benzoic acid intermediate and sulfuric acid described in step 3) is preferably 14 - 17 mmol:12 mL, more preferably 15 - 16 mmol:12 mL, and even more preferably 15.3 mmol:12 mL; the mass concentration of sulfuric acid is preferably 92 - 98%, more preferably 94 - 96%; the temperature of the ring-forming reaction is preferably 125 - 145 °C, more preferably 130 - 140 °C, and even more preferably 135 °C; the time of the ring-forming reaction is preferably 2 - 4 h, more preferably 2.5 - 3.5 h, and even more preferably 3 h.

[0052] In the present invention, the cyclization reaction in step 3) is preferably carried out under stirring conditions, and the stirring speed is preferably 400 - 600 rpm, more preferably 500 rpm.

[0053] In the present invention, after the cyclization reaction in step 3) is completed, the reaction solution is preferably placed in ice water to obtain a crude mixture, and the crude mixture is successively subjected to recrystallization, filtration, and drying to obtain acridone intermediate 1; the temperature of the ice water is preferably -10 to 10 °C, more preferably 0 °C, and the volume ratio of ice water to sulfuric acid is preferably 100:10 - 15, more preferably 100:12; the recrystallization is preferably carried out by adjusting the pH value of the crude mixture to 12 - 13 with a saturated sodium hydroxide solution; the drying temperature is preferably 40 - 60 °C, more preferably 50 °C; the drying time is preferably 20 - 28 h, more preferably 24 h.

[0054] In the present invention, the acridone intermediate 1 in step 3) is 3,6-diamino-9(10H)-acridone, and its structural formula is:

[0055] In the present invention, the molar volume ratio of the acridone intermediate 1 to butyric anhydride in step 4) is preferably 0.8 - 2.5 mmol:10 mL, more preferably 1 - 2 mmol:10 mL, and even more preferably 1.33 mmol:10 mL; the temperature of the condensation reaction is preferably 90 - 110 °C, more preferably 95 - 105 °C, and even more preferably 100 °C; the condensation reaction time is preferably 1 - 3 h, more preferably 1.5 - 2.5 h, and even more preferably 2 h.

[0056] In the present invention, the condensation reaction in step 4) is preferably carried out under stirring conditions, and the stirring speed is preferably 400 - 600 rpm, more preferably 500 rpm.

[0057] In the present invention, after the condensation reaction in step 4) is completed, it is preferably successively subjected to recrystallization, filtration, and drying to obtain acridone intermediate 2; the recrystallization is preferably carried out by adjusting the pH value of the reaction solution to 12 - 13 with a sodium hydroxide solution, and the mass concentration of the sodium hydroxide solution is preferably 37 - 43%, more preferably 40%; the drying temperature is preferably 40 - 60 °C ， more preferably 50 °C; the drying time is preferably 20 - 28 h, more preferably 24 h.

[0058] In the present invention, the acridone intermediate 2 in step 4) is 3,6-bis(butyrylamino)-9(10H)-acridone, and its structural formula is:

[0059] In the present invention, the molar volume ratio of the acridone intermediate 2 to phosphorus oxychloride in step 5) is preferably 12-15 mmol: 20 mL, more preferably 13-14 mmol: 20 mL, and even more preferably 13.7 mmol: 20 mL; the temperature of the electrophilic substitution reaction is preferably 80-100 °C, more preferably 85-95 °C, and even more preferably 90 °C; the time of the electrophilic substitution reaction is preferably 1-2 h, more preferably 1.5 h.

[0060] In the present invention, the electrophilic substitution reaction in step 5) is preferably carried out under stirring conditions, and the stirring speed is preferably 400-600 rpm, more preferably 500 rpm.

[0061] In the present invention, after the electrophilic substitution reaction in step 5) is completed, the reaction solution is preferably cooled to room temperature, and then subjected to reduced pressure distillation, neutralization of phosphorus oxychloride, extraction to obtain an organic layer, and the organic layer is successively dried, subjected to reduced pressure distillation, and silica gel column chromatography to obtain an acridine intermediate; the temperature of the reduced pressure distillation is independently preferably 50-60 °C, more preferably 55 °C; the pressure of the reduced pressure distillation is independently preferably 0.05-0.15 MPa, more preferably 0.1 MPa; the time of the reduced pressure distillation is independently preferably 5-15 min, more preferably 10 min; the neutralization of phosphorus oxychloride is preferably to place the product of the reduced pressure distillation in ice water to obtain a crude product, and the pH value of the crude product is adjusted to 12-13 with a saturated sodium hydroxide solution; the temperature of the ice water is preferably -10-10 °C, more preferably 0 °C, and the volume ratio of the ice water to phosphorus oxychloride is preferably 100: 15-25, more preferably 100: 20; the extraction reagent is preferably dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is preferably 9-11: 1, more preferably 10: 1; the number of extraction times is preferably 2-4 times, more preferably 3 times; the drying reagent is preferably anhydrous magnesium sulfate; the silica gel column chromatography reagent is preferably dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is preferably 20-40: 1, more preferably 30: 1. The purpose of the first reduced pressure distillation is to remove the solvent phosphorus oxychloride, and the purpose of the second reduced pressure distillation is to remove dichloromethane and methanol.

[0062] In the present invention, the acridine intermediate in step 5) is 3,6-bis(butyramido)-9-chloroacridine, and the structural formula is:

[0063] In the present invention, the molar volume ratio of the acridine intermediate, N,N-dimethyl-p-phenylenediamine and phenol described in step 6) is preferably 1 mmol: 3-5 mmol: 2-4 mL, more preferably 1 mmol: 3.5-4.5 mmol: 2.5-3.5 mL, and even more preferably 1 mmol: 4 mmol: 2.88 mL; the temperature of the nucleophilic substitution reaction is preferably 90-110 °C, more preferably 95-105 °C, and even more preferably 100 °C; the time of the nucleophilic substitution reaction is preferably 1-3 h, more preferably 1.5-2.5 h, and even more preferably 2 h.

[0064] In the present invention, the nucleophilic substitution reaction described in step 6) is preferably carried out under stirring conditions, and the stirring speed is preferably 400-600 rpm, more preferably 500 rpm.

[0065] In the present invention, after the nucleophilic substitution reaction described in step 6) is completed, a quenching reaction and extraction are preferably carried out in sequence to obtain an organic layer, and the organic layer is sequentially dried, subjected to vacuum distillation, and silica gel column chromatography to obtain the trisubstituted acridine intermediate 1; the reagent used for the quenching reaction is preferably water, and the volume ratio of water to phenol is preferably 100: 10-15, more preferably 100: 12; the reagents used for the extraction are preferably dichloromethane and saturated sodium chloride solution, and the volume ratio of dichloromethane and saturated sodium chloride solution is preferably 1-2: 1, more preferably 1.5: 1; the number of extraction times is preferably 2-4 times, more preferably 3 times; the reagent used for drying is preferably anhydrous magnesium sulfate; the temperature of the vacuum distillation is preferably 50-60 °C, more preferably 55 °C; the pressure of the vacuum distillation is preferably 0.05-0.15 MPa, more preferably 0.1 MPa; the time of the vacuum distillation is preferably 10-20 min, more preferably 15 min; the reagents used for the silica gel column chromatography are preferably dichloromethane and methanol, and the volume ratio of dichloromethane and methanol is preferably 10-30: 1, more preferably 20: 1. The purpose of vacuum distillation is to remove dichloromethane and methanol.

[0066] In the present invention, the trisubstituted acridine intermediate 1 described in step 6) is 3,6-bis(butyrylamino)-9-[4'-(N,N-dimethylamino)phenylamino]acridine, and the structural formula is:

[0067] In the present invention, the molar volume ratio of the trisubstituted acridine intermediate 1 to sulfuric acid in step 7) is preferably 2 - 4 mmol: 10 mL, more preferably 2.68 - 3.5 mmol: 10 mL, and still more preferably 3 mmol: 10 mL; the mass concentration of sulfuric acid is preferably 92 - 98%, more preferably 94 - 96%; the temperature of the hydrolysis reaction is preferably 90 - 110 °C, more preferably 95 - 105 °C, and still more preferably 100 °C; the time of the hydrolysis reaction is preferably 20 - 40 min, more preferably 25 - 35 min, and still more preferably 30 min.

[0068] In the present invention, the hydrolysis reaction in step 7) is preferably carried out under stirring conditions, and the stirring speed is preferably 400 - 600 rpm, more preferably 500 rpm.

[0069] In the present invention, after the hydrolysis reaction in step 7) is completed, the reaction solution is preferably cooled to room temperature, and then successively neutralized with sulfuric acid, recrystallized, filtered, and dried to obtain the trisubstituted acridine intermediate 2; the neutralization of sulfuric acid is preferably to place the reaction solution in ice water to obtain a crude mixture; the temperature of the ice water is preferably -10 - 10 °C, more preferably 0 °C, and the volume ratio of ice water to sulfuric acid is preferably 50:8 - 12, more preferably 50:10; the recrystallization is preferably to adjust the pH value of the crude mixture to 12 - 13 with a saturated sodium hydroxide solution; the drying temperature is preferably 40 - 50 °C, more preferably 45 °C; the drying time is preferably 20 - 28 h, more preferably 24 h.

[0070] In the present invention, the trisubstituted acridine intermediate 2 in step 7) is 3,6-diamino-9-[4'-(N,N-dimethylamino)phenylamino]acridine, and its structural formula is:

[0071] In the present invention, the molar volume ratio of the trisubstituted acridine intermediate 2, 3-chloropropionyl chloride, triethylamine, and N,N-dimethylformamide in step 8) is preferably 1 mmol: 4 - 6 mmol: 0.3 - 0.6 mL: 1 - 3 mL, more preferably 1 mmol: 4.5 - 5.5 mmol: 0.4 - 0.5 mL: 1.67 - 2.5 mL, and still more preferably 1 mmol: 5 mmol: 0.42 mL: 2 mL; the temperature of the nucleophilic addition reaction is preferably 50 - 70 °C, more preferably 55 - 65 °C, and still more preferably 60 °C; the time of the nucleophilic addition reaction is preferably 1 - 3 h, more preferably 1.5 - 2.5 h, and still more preferably 2 h.

[0072] In the present invention, the nucleophilic addition reaction in step 8) is preferably carried out under stirring conditions, and the stirring speed is preferably 400 - 600 rpm, more preferably 500 rpm.

[0073] In the present invention, in step 8), the mixing preferably first mixes the trisubstituted acridine intermediate 2 and triethylamine, and then successively adds N,N-dimethylformamide and 3-chloropropionyl chloride.

[0074] In the present invention, after the nucleophilic addition reaction in step 8), it is preferably followed by a quenching reaction, distillation under reduced pressure, extraction to obtain an organic layer, and the organic layer is successively dried, distilled under reduced pressure, and subjected to silica gel column chromatography to obtain the trisubstituted acridine intermediate; the reagent used for the quenching reaction is water, and the volume ratio of water to N,N-dimethylformamide is preferably 20-30:1, more preferably 25:1; the temperature of the distillation under reduced pressure is independently preferably 60-70°C, more preferably 65°C; the pressure of the distillation under reduced pressure is independently preferably 0.05-0.15 MPa, more preferably 0.1 MPa; the time of the distillation under reduced pressure is independently preferably 20-30 min, more preferably 25 min; the reagents used for the extraction are preferably dichloromethane and saturated sodium chloride solution, and the volume ratio of dichloromethane to saturated sodium chloride solution is preferably 1-2:1, more preferably 1.5:1; the number of extractions is preferably 2-4 times, more preferably 3 times; the reagent used for the drying is preferably anhydrous magnesium sulfate; the reagents used for the silica gel column chromatography are preferably dichloromethane and methanol, and the volume ratio of dichloromethane to methanol is preferably 10-30:1, more preferably 20:1.

[0075] The present invention also provides the use of the trisubstituted acridine derivative containing a piperazine group in the preparation of anti-tumor drugs and anti-inflammatory drugs.

[0076] In the present invention, the anti-tumor drugs and anti-inflammatory drugs are independently preferably injection agents, tablets, capsules, aerosols, suppositories, membranes, dripping pills, ointments, controlled release agents, sustained release agents or nano-formulations.

[0077] The technical solutions provided by the present invention are described in detail below with reference to the examples, but they should not be construed as limiting the protection scope of the present invention.

[0078] Example 1

[0079] Mix 116.6 mmol of 2-chloro-4-aminobenzoic acid and 30 mL of acetic anhydride, and carry out a condensation reaction by stirring at 100°C at a speed of 800 rpm for 30 min to obtain a clear solution. Add 90 mL of dichloromethane to the clear solution to precipitate, and then filter with a sintered glass funnel to obtain a solid product. Wash the solid product 3 times with dichloromethane to obtain the condensation intermediate: 4-(acetylamino)-2-chlorobenzoic acid. The condensation intermediate is a white solid with a mass of 17.2 g and a yield of 69.1%.

[0080] 2.34 mmol of anhydrous potassium carbonate, 0.46 mmol of copper powder, 0.43 mmol of cuprous oxide, 4.68 mmol of condensation intermediate, 5.15 mmol of m-aminophenacetanilide and 8 mL of ethylene glycol monoethyl ether were mixed and stirred at 135 °C at a speed of 500 rpm for 5 h to carry out the Ullmann reaction to obtain a reaction mixture. The reaction mixture was poured into 30 mL of distilled water and filtered through a diatomaceous earth filter to obtain a filtrate. Hydrochloric acid (the volume ratio of HCl to water in hydrochloric acid was 3:1) was added to the filtrate for acidification, and the acidification was terminated when the pH value of the filtrate reached 2.5. Filtration was carried out again to obtain a crude product. The crude product was dried at 50 °C for 24 h to obtain a benzoic acid intermediate: 4-(acetamido)-2-(3-(acetamido)phenylamino)benzoic acid. The benzoic acid intermediate was a gray solid with a mass of 420 mg and a yield of 27.4%.

[0081] Sulfuric acid with a mass concentration of 96% and a volume of 12 mL was added dropwise to 15.3 mmol of the benzoic acid intermediate at a rate of 1 drop / s, and the mixture was stirred at 135 °C at a speed of 500 rpm for 2 h to carry out the cyclization reaction. After the reaction was completed, it was cooled to room temperature to obtain a reaction mixture. The reaction mixture was poured into 100 mL of ice water at 0 °C to obtain a crude mixture, and the pH value of the crude mixture was adjusted to 12.5 with saturated sodium hydroxide solution, and then filtered through a fritted funnel to obtain a filter residue. The filter residue was dried at 45 °C for 24 h to obtain an acridone intermediate 1: 3,6-diamino-9(10H)-acridone. The acridone intermediate 1 was a yellowish-green solid and was directly used in the next reaction step without further purification.

[0082] 1.33 mmol of the acridone intermediate 1 was placed in a 100 mL round-bottom flask, 10 mL of butyric anhydride was added, and the mixture was stirred at 100 °C at a speed of 500 rpm for 2 h to carry out the condensation reaction to obtain a reaction mixture. The pH value of the reaction mixture was adjusted to 12.5 with a sodium hydroxide solution with a mass concentration of 40%, and then filtered through a fritted funnel to obtain a filter residue. The filter residue was dried at 50 °C for 24 h to obtain an acridone intermediate 2: 3,6-bis(butyramido)-9(10H)-acridone. The acridone intermediate 2 was a gray solid and was directly used in the next reaction step without further purification.

[0083] To 13.7 mmol of acridone intermediate 2, 20 mL of phosphorus oxychloride was added, and an electrophilic substitution reaction was carried out by stirring at 90 °C at a rotation speed of 500 rpm for 1.5 h. After the reaction was completed, it was cooled to room temperature to obtain a reaction mixture. The reaction mixture was placed in a rotary evaporator and distilled under reduced pressure at 55 °C and 0.1 MPa for 10 min. The product of the reduced pressure distillation was poured into 100 mL of ice water at 0 °C to obtain a crude product, and the pH value of the crude product was adjusted to 12.5 with saturated sodium hydroxide solution to obtain a mixed solution. The mixed solution was extracted with dichloromethane and methanol with a volume ratio of 10:1 for 3 times to obtain an organic layer. The organic layer was dried with anhydrous magnesium sulfate and then placed in a rotary evaporator and distilled under reduced pressure at 55 °C and 0.1 MPa for 15 min. The product of the reduced pressure distillation was subjected to silica gel column chromatography with dichloromethane and methanol with a volume ratio of 30:1 (the dropping rate of the product of the reduced pressure distillation was 5 drops / s) to obtain an acridine intermediate: 3,6-bis(butyrylamino)-9-chloroacridine. The acridine intermediate was a bright yellow solid with a mass of 1.6 g and a yield of 30.5%.

[0084] 4.16 mmol of acridine intermediate, 16.66 mmol of N,N-dimethyl-p-phenylenediamine and 12 mL of phenol were mixed, and a nucleophilic substitution reaction was carried out by stirring at 100 °C at a rotation speed of 500 rpm for 2 h to obtain a reaction mixture. The reaction mixture was poured into 100 mL of distilled water, and the pH value of the reaction mixture was adjusted to 12.5 with saturated sodium hydroxide solution. Then, it was extracted with dichloromethane and saturated sodium chloride solution with a volume ratio of 1:1 for 3 times to obtain an organic layer. The organic layer was dried with anhydrous magnesium sulfate and then placed in a rotary evaporator and distilled under reduced pressure at 55 °C and 0.1 MPa for 15 min. The product of the reduced pressure distillation was subjected to silica gel column chromatography with dichloromethane and methanol with a volume ratio of 20:1 (the dropping rate of the product of the reduced pressure distillation was 5 drops / s) to obtain a trisubstituted acridine intermediate 1: 3,6-bis(butyrylamino)-9-[4'-(N,N-dimethylamino)phenylamino]acridine. The trisubstituted acridine intermediate 1 was a red-brown solid with a mass of 1.3 g and a yield of 64.7%.

[0085] 10 mL of sulfuric acid with a mass concentration of 96% was added dropwise to 2.68 mmol of trisubstituted acridine intermediate 1 at a rate of 1 drop / s, and a hydrolysis reaction was carried out by stirring at 100 °C at a rotation speed of 500 rpm for 0.5 h. After the reaction was completed, it was cooled to room temperature to obtain a reaction mixture. The reaction mixture was poured into 50 mL of ice water at 0 °C to obtain a crude product, and the pH value of the crude product was adjusted to 12.5 with saturated sodium hydroxide solution. Then, it was filtered through a sintered glass funnel to obtain a filter residue, and the filter residue was dried at 45 °C for 24 h to obtain a trisubstituted acridine intermediate 2: 3,6-diamino-9-[4'-(N,N-dimethylamino)phenylamino]acridine. The trisubstituted acridine intermediate 2 was a red-brown solid and was directly used in the next reaction step without further purification.

[0086] Mix 2.39 mmol of the trisubstituted acridine intermediate 2 and 1 mL of triethylamine, then successively add 4 mL of N,N-dimethylformamide and 11.95 mmol of 3-chloropropionyl chloride, and place it at 60 °C and stir at a speed of 500 rpm for 2 h to carry out a nucleophilic addition reaction to obtain a reaction mixture. Pour the reaction mixture into 100 mL of distilled water, and place it in a rotary evaporator. Distill it under reduced pressure at 65 °C and 0.1 MPa for 25 min. Extract the product obtained by reduced pressure distillation 3 times with a dichloromethane and saturated sodium chloride solution with a volume ratio of 1:1 to obtain an organic layer. Dry the organic layer with anhydrous magnesium sulfate and then place it in a rotary evaporator. Distill it under reduced pressure at 55 °C and 0.1 MPa for 15 min. Perform silica gel column chromatography on the product obtained by reduced pressure distillation with a dichloromethane and methanol with a volume ratio of 20:1 (the dropping rate of the product obtained by reduced pressure distillation is 5 drops / s) to obtain the trisubstituted acridine intermediate: N,N'-(9-(4-(dimethylamino)phenylamino)acridine-3,6-diyl)bis(3-chloropropanamide). The trisubstituted acridine intermediate is a red-brown solid with a mass of 550 mg and a yield of 44.7%.

[0087] Mix 1.05 mmol of the trisubstituted acridine intermediate, 0.5 mmol of potassium iodide and 0.5 mmol of potassium carbonate, then successively add 15 mL of absolute ethanol and 5.24 mmol of N-methylpiperazine, and place it at 90 °C and stir at a speed of 500 rpm for 2 h to carry out a nucleophilic substitution reaction to obtain a reaction mixture. Pour the reaction mixture into 400 mL of distilled water, and place it in a rotary evaporator. Distill it under reduced pressure at 50 °C and 0.1 MPa for 25 min. Extract the product obtained by reduced pressure distillation 3 times with a dichloromethane and saturated sodium chloride solution with a volume ratio of 1:1 to obtain an organic layer. Dry the organic layer with anhydrous magnesium sulfate and then place it in a rotary evaporator. Distill it under reduced pressure at 50 °C and 0.1 MPa for 25 min. Perform silica gel column chromatography on the product obtained by reduced pressure distillation with a dichloromethane and methanol with a volume ratio of 20:1 (the dropping rate of the product obtained by reduced pressure distillation is 5 drops / s) to obtain the trisubstituted acridine derivative containing a piperazine group: N,N'-(9-((4-(dimethylamino)phenyl)amino)acridine-3,6-diyl)bis(3-(4-methylpiperazin-1-yl)propanamide). The trisubstituted acridine derivative containing a piperazine group is a red-brown solid with a mass of 60 mg and a yield of 8.87%.

[0088] Perform NMR characterization on the trisubstituted acridine derivative containing a piperazine group prepared in Example 1. Figure 1 For the trisubstituted acridine derivative containing a piperazine group prepared in Example 1 1 HNMR nuclear magnetic resonance spectrum.

[0089] Figure 1 The obtained 1The \(^1H\) NMR is as follows: 1 \(^1H\) NMR (400 MHz, Methanol-d4) δ 8.42 (d, J = 2.1 Hz, 2H), 8.01 (d, J = 9.4 Hz, 2H), 7.24–7.17 (m, 4H), 6.85 (d, J = 8.8 Hz, 2H), 3.03 (s, 6H), 2.83 (t, J = 7.0 Hz, 5H), 2.78–2.39 (m, 20H), 2.36 (s, 6H).

[0090] It can be seen from Figure 1 that the trisubstituted acridine derivative containing a piperazine group prepared in Example 1 of the present invention is N,N'-(9-((4-(dimethylamino)phenyl)amino)acridine-3,6-diyl)bis(3-(4-methylpiperazin-1-yl)propanamide).

[0091] Example 2

[0092] 110 mmol of 2-chloro-4-aminobenzoic acid and 30 mL of acetic anhydride were mixed and stirred at 110 °C at a speed of 800 rpm for 20 min for a condensation reaction to obtain a clear solution. 90 mL of dichloromethane was added to the clear solution to precipitate a solid, and then the solid product was filtered through a sintered glass funnel. The solid product was washed 3 times with dichloromethane to obtain a condensation intermediate: 4-(acetamido)-2-chlorobenzoic acid.

[0093] 3.74 mmol of anhydrous potassium carbonate, 0.56 mmol of copper powder, 0.56 mmol of cuprous oxide, 4.68 mmol of the condensation intermediate, 3.75 mmol of m-aminoacetanilide and 9.5 mL of ethylene glycol monoethyl ether were mixed and stirred at 145 °C at a speed of 500 rpm for 4 h for an Ullmann reaction to obtain a reaction mixture. The reaction mixture was poured into 38 mL of distilled water and filtered through a diatomaceous earth filter to obtain a filtrate. Hydrochloric acid (the volume ratio of HCl to water in the hydrochloric acid is 3:1) was added to the filtrate for acidification, and the acidification was terminated when the pH value of the filtrate reached 2.5. The crude product was obtained by filtration again. The crude product was dried at 50 °C for 24 h to obtain a benzoic acid intermediate: 4-(acetamido)-2-(3-(acetamido)phenylamino)benzoic acid.

[0094] Add sulfuric acid with a mass concentration of 92% and a volume of 12 mL dropwise to 17 mmol of benzoic acid intermediate at a rate of 1 drop / s, and place it at 125 °C and stir at a speed of 500 rpm for 4 h for cyclization reaction. After the reaction is completed, cool it to room temperature to obtain a reaction mixture. Pour the reaction mixture into 100 mL of ice water at 0 °C to obtain a crude mixture, adjust the pH value of the crude mixture to 12 with saturated sodium hydroxide solution, then filter it through a sintered funnel to obtain a filter residue, and dry the filter residue at 50 °C for 24 h to obtain acridone intermediate 1: 3,6-diamino-9(10H)-acridone. Acridone intermediate 1 is a yellowish-green solid and can be directly used in the next reaction step without further purification.

[0095] Place 2 mmol of acridone intermediate 1 in a 100 mL round-bottom flask, add 10 mL of butyric anhydride, and stir at 90 °C at a speed of 500 rpm for 3 h for condensation reaction to obtain a reaction mixture. Adjust the pH value of the reaction mixture to 13 with sodium hydroxide solution with a mass concentration of 43%, then filter it through a sintered funnel to obtain a filter residue, and dry the filter residue at 50 °C for 24 h to obtain acridone intermediate 2: 3,6-bis(butyrylamino)-9(10H)-acridone. Acridone intermediate 2 is a gray solid and can be directly used in the next reaction step without further purification.

[0096] Add 20 mL of phosphorus oxychloride to 15 mmol of acridone intermediate 2, and stir at 100 °C at a speed of 400 rpm for 1 h for electrophilic substitution reaction. After the reaction is completed, cool it to room temperature to obtain a reaction mixture. Place the reaction mixture in a rotary evaporator and distill it under reduced pressure at 55 °C and 0.1 MPa for 10 min. Pour the product of reduced pressure distillation into 80 mL of ice water at 0 °C to obtain a crude product, and adjust the pH value of the crude product to 12 with saturated sodium hydroxide solution to obtain a mixed solution. Extract the mixed solution with dichloromethane and methanol with a volume ratio of 9:1 for 3 times to obtain an organic layer. Dry the organic layer with anhydrous magnesium sulfate and then place it in a rotary evaporator and distill it under reduced pressure at 55 °C and 0.1 MPa for 15 min. Perform silica gel column chromatography on the product of reduced pressure distillation with dichloromethane and methanol with a volume ratio of 40:1 (the dropping rate of the product of reduced pressure distillation is 5 drops / s) to obtain acridine intermediate: 3,6-bis(butyrylamino)-9-chloroacridine. The acridine intermediate is a bright yellow solid.

[0097] 4.16 mmol of acridine intermediate, 20 mmol of N,N-dimethyl-p-phenylenediamine and 16.5 mL of phenol were mixed and stirred at 500 rpm for 3 h at 90 °C for nucleophilic substitution reaction to obtain a reaction mixture. The reaction mixture was poured into 110 mL of distilled water, and the pH value of the reaction mixture was adjusted to 12 with saturated sodium hydroxide solution. Then, it was extracted 3 times with a 1:1 volume ratio of dichloromethane and saturated sodium chloride solution to obtain an organic layer. The organic layer was dried with anhydrous magnesium sulfate and then placed in a rotary evaporator, and vacuum distilled at 55 °C and 0.1 MPa for 15 min. The product of vacuum distillation was subjected to silica gel column chromatography with a 10:1 volume ratio of dichloromethane and methanol (the dropping rate of the product of vacuum distillation was 5 drops / s) to obtain trisubstituted acridine intermediate 1: 3,6-bis(butyrylamino)-9-[4'-(N,N-dimethylamino)phenylamino]acridine. Trisubstituted acridine intermediate 1 was a red-brown solid.

[0098] 10 mL of sulfuric acid with a mass concentration of 92% was added dropwise to 4 mmol of trisubstituted acridine intermediate 1 at a rate of 1 drop / s, and it was stirred at 500 rpm at 90 °C for 40 min for hydrolysis reaction. After the reaction, it was cooled to room temperature to obtain a reaction mixture. The reaction mixture was poured into 42 mL of ice water at 0 °C to obtain a crude product. The pH value of the crude product was adjusted to 13 with saturated sodium hydroxide solution, and then filtered through a sintered glass funnel to obtain a filter residue. The filter residue was dried at 45 °C for 24 h to obtain trisubstituted acridine intermediate 2: 3,6-diamino-9-[4'-(N,N-dimethylamino)phenylamino]acridine. Trisubstituted acridine intermediate 2 was a red-brown solid and was directly used in the next reaction step without further purification.

[0099] 2.39 mmol of trisubstituted acridine intermediate 2 and 1.4 mL of triethylamine were mixed, and then 7 mL of N,N-dimethylformamide and 14 mmol of 3-chloropropionyl chloride were successively added. It was stirred at 500 rpm at 70 °C for 2 h for nucleophilic addition reaction to obtain a reaction mixture. The reaction mixture was poured into 150 mL of distilled water and placed in a rotary evaporator, and vacuum distilled at 65 °C and 0.1 MPa for 25 min. The product of vacuum distillation was extracted 3 times with a 1:1 volume ratio of dichloromethane and saturated sodium chloride solution to obtain an organic layer. The organic layer was dried with anhydrous magnesium sulfate and then placed in a rotary evaporator, and vacuum distilled at 55 °C and 0.1 MPa for 15 min. The product of vacuum distillation was subjected to silica gel column chromatography with a 30:1 volume ratio of dichloromethane and methanol (the dropping rate of the product of vacuum distillation was 5 drops / s) to obtain trisubstituted acridine intermediate: N,N'-(9-(4-(dimethylamino)phenylamino)acridine-3,6-diyl)bis(3-chloropropionamide). The trisubstituted acridine intermediate was a red-brown solid.

[0100] 1.05 mmol of the trisubstituted acridine intermediate, 0.42 mmol of potassium iodide and 0.63 mmol of potassium carbonate were mixed, and then 18 mL of absolute ethanol and 4.2 mmol of N-methylpiperazine were sequentially added. The mixture was placed at 80 °C and stirred at a speed of 400 rpm for 3 h for a nucleophilic substitution reaction to obtain a reaction mixture. The reaction mixture was poured into 400 mL of distilled water and placed in a rotary evaporator. It was distilled under reduced pressure at 45 °C and 0.15 MPa for 30 min. The product of the reduced pressure distillation was extracted 3 times with a volume ratio of 2:1 dichloromethane and saturated sodium chloride solution to obtain an organic layer. The organic layer was dried with anhydrous magnesium sulfate and then placed in a rotary evaporator. It was distilled under reduced pressure at 45 °C and 0.15 MPa for 20 min. The product of the reduced pressure distillation was subjected to silica gel column chromatography with a volume ratio of 10:1 dichloromethane and methanol (the dropping rate of the product of the reduced pressure distillation was 5 drops / s) to obtain a trisubstituted acridine derivative containing a piperazine group: N,N'-(9-((4-(dimethylamino)phenyl)amino)acridine-3,6-diyl)bis(3-(4-methylpiperazin-1-yl)propanamide). The trisubstituted acridine derivative containing a piperazine group was a red-brown solid.

[0101] The MTT method was used to detect the antitumor activity of the trisubstituted acridine derivative containing a piperazine group prepared in Example 1. The detection steps were as follows:

[0102] The ovarian cancer cells ES2 in the logarithmic growth phase were inoculated into 96-well plates (inoculation density: 5000 cells / well, 160 μL of medium / well, the medium was high-glucose DMEM medium, the medium brand was GIBCO, product number was C11965500BT, packaging specification was 500 mL), and the 96-well plates were placed in an incubator at 37 °C and 5% CO2 for 12 h. Take out the 96-well plates, divide the 96-well plates into two groups of test wells, namely the control group and the test group, and each group of tests contains 3 wells. Add 40 μL / well of dimethyl sulfoxide medium (the mass concentration of dimethyl sulfoxide in the medium was 0.1%, the medium was high-glucose DMEM medium, the medium brand was GIBCO, product number was C11965500BT, packaging specification was 500 mL) to the wells of the control group; add 40 μL / well of the three-substituted acridine derivative medium containing a piperazine group (the concentration of the three-substituted acridine derivative containing a piperazine group in the medium was 10 μM, the medium was high-glucose DMEM medium, the medium brand was GIBCO, product number was C11965500BT, packaging specification was 500 mL) to the wells of the test group. Place the 96-well plates in an incubator at 37 °C and 5% CO2 for 48 h. Take out the 96-well plates, add 20 μL of MTT working solution with a concentration of 5 mg / mL (i.e., thiazolyl blue working solution, using physiological saline as the solvent) to each well, and then place it in an incubator at 37 °C and 5% CO2 for continued culture for 4 h. After the culture is completed, under light-proof conditions, use a vacuum pump to suck out the supernatant until there is no liquid residue in the wells, add 100 μL of dimethyl sulfoxide to each well, and shake it in the dark at room temperature for 10 min to ensure that the purple formazan crystals are completely dissolved. Measure the absorbance value of the solution in each well at 570 nm to obtain the ovarian cancer cell viability level OD. According to the formula: cell survival rate = OD 试验组 / OD 对照组 × 100%, calculate the cell survival rate.

[0103] Repeat the above detection steps for breast cancer cells MDA-MB-231, colorectal cancer cells HCT116, human non-small cell lung cancer cells H1581, liver cancer cells Huh-7, and human glioma cells U87 in the logarithmic growth phase respectively, and obtain the inhibition rates of the three-substituted acridine derivative containing a piperazine group on different tumor cells.

[0104] Figure 2 The inhibitory activity of the three-substituted acridine derivative containing a piperazine group prepared in Example 1 on tumor cells. By Figure 2It can be seen that the trisubstituted acridine derivative containing a piperazine group of the present invention has good anti-tumor activity against breast cancer cells MDA-MB-231, colorectal cancer cells HCT116, human non-small cell lung cancer cells H1581, liver cancer cells Huh-7, human glioma cells U87 and ovarian cancer cells ES2. The inhibition rates against ovarian cancer cells ES2 and human glioma cells U87 reach 20%, the inhibition rates against colorectal cancer cells HCT116 and liver cancer cells Huh-7 reach 30%, the inhibition rate against breast cancer cells MDA-MB-231 reaches 30%, and the inhibition rate against lung cancer cells H1581 reaches 40%. This result indicates that the trisubstituted acridine derivative containing a piperazine group of the present invention has broad-spectrum anti-tumor activity.

[0105] By establishing an LPS-induced RAW264.7 cell inflammation model, the inhibitory effect of the trisubstituted acridine derivative containing a piperazine group prepared in Example 1 on the secretion of TNF-α and IL-6 was detected. The detection steps are as follows:

[0106] Mouse macrophages RAW264.7 were inoculated into 96-well plates (inoculation density: 100,000 cells / well, 160 μL of medium / well, the medium was high-glucose medium DMEM, the medium brand was GIBCO, the product number was C11965500BT, and the packaging specification was 500 mL). The 96-well plates were placed in an incubator at 37 °C and 5% CO2 for overnight culture to ensure that the cells adhered completely. The 96-well plates were taken out and divided into three groups of test wells, with 3 wells in each group. In two groups of test wells, 40 μL of the medium containing the trisubstituted acridine derivative with a piperazine group (the concentration of the trisubstituted acridine derivative with a piperazine group in the medium was 10 μmol / L, the medium was high-glucose medium DMEM, the medium brand was GIBCO, the product number was C11965500BT, and the packaging specification was 500 mL) was added; the other group was not treated and served as the blank group. Then, a DMEM solution of LPS (lipopolysaccharide) with a concentration of 10.5 ng / μL was added to each well to make the concentration of LPS in the well reach 0.5 ng / μL. The 96-well plates were placed in an incubator at 37 °C and 5% CO2 for 12 h. The 96-well plates were taken out, and the cell supernatants in the wells were collected. The contents of TNF-α and IL-6 in the cell supernatants were detected using a murine TNF-α kit and an IL-6 ELISA kit (both kits were purchased from Shanghai Beyotime Biotechnology Co., Ltd.). Based on the blank group, the inhibition rates of TNF-α and IL-6 secretion were calculated.

[0107] Figure 3 For the inhibitory activity of the trisubstituted acridine derivative containing a piperazine group prepared in Example 1 on inflammatory factors. By Figure 3It can be seen that the trisubstituted acridine derivative containing a piperazine group of the present invention can effectively inhibit the secretion of inflammatory factors in LPS-induced RAW264.7 cells, with the inhibitory effect on TNF-α secretion reaching 15% and the inhibitory effect on IL-6 secretion reaching 20%.

[0108] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements can be made, and these improvements and refinements should also be regarded as the protection scope of the present invention.

Claims

1. A trisubstituted acridine derivative containing a piperazine group, characterized in that: The structural formula of the trisubstituted acridine derivative containing a piperazine group is:

2. The method for preparing a trisubstituted acridine derivative containing a piperazine group according to claim 1, characterized in that: The following steps are included: The trisubstituted acridine intermediate, N-methylpiperazine, potassium iodide, potassium carbonate and anhydrous ethanol are mixed and subjected to a nucleophilic substitution reaction to obtain a trisubstituted acridine derivative containing a piperazine group.

3. The method for preparing a trisubstituted acridine derivative containing a piperazine group according to claim 2, characterized in that: The structural formula of the trisubstituted acridine intermediate is:

4. The method for preparing a trisubstituted acridine derivative containing a piperazine group according to claim 2 or 3, characterized in that: The molar ratio of the trisubstituted acridine intermediate, N-methylpiperazine, potassium iodide and potassium carbonate is 1:4-6:0.4-0.6:0.4-0.6, and the molar volume ratio of the trisubstituted acridine intermediate and anhydrous ethanol is 1mmol:13-18mL.

5. The method for preparing a trisubstituted acridine derivative containing a piperazine group according to claim 4, characterized in that: The temperature of the nucleophilic substitution reaction is 80-100° C., and the time of the nucleophilic substitution reaction is 1-3 hours.

6. The method for preparing a trisubstituted acridine derivative containing a piperazine group according to claim 5, characterized in that: The nucleophilic substitution reaction is carried out under stirring conditions, and the stirring speed is 400-600 rpm.

7. Use of the trisubstituted acridine derivative containing a piperazine group according to claim 1 in the preparation of antitumor drugs and anti-inflammatory drugs.

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